Pharmacological activities of Artemisia absinthium and control of hepatic cancer by expression regulation of TGFß1 and MYC genes.

Plant derived compounds have always been an important source of medicines and have received significant attention in recent years due to their diverse pharmacological properties. Millions of plant-based herbal or traditional medicines are used to cure various types of cancers especially due to activation of proliferative genes. The aim of the present study was to characterize the altered and attenuated gene expression of the selected growth factor namely Transforming growth factor Beta -1 (TGFß1) and MYC in human hepatoma-derived (Huh7) liver cancer cell lines in response to extracts of Artemisia absinthium dissolved in selected organic solvents. Ethanolic, methanolic and acetone extract of different plant parts (leaf, stem and flowers) was used to access the antiproliferative activity by MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide) assay. Quantitative Real-Time RT-PCR revealed that the transcript levels of TGFß1 are induced in the samples treated with methanolic extract of Artemisia absinthium. Furthermore, reduced expression levels of MYC gene was noticed in cancerous cells suggesting antiproliferative properties of the plant. This study further highlights the resistance profile of various microbes by antimicrobial susceptibility test with plant extracts. In addition, antidiabetic effect of Artemisia absinthium have also shown positive results. Our study elucidates the potentials of Artemisia absinthium as a medicinal plant, and highlights the differential expression of genes involved in its mitogenic and anti-proliferative activity with a brief account of its pharmacological action.

Effects of Drying Temperature and Solvents on In Vitro Diabetic Wound Healing Potential of Moringa oleifera Leaf Extracts.

The delayed healing of wounds among people with diabetes is a severe problem worldwide. Hyperglycemia and increased levels of free radicals are the major inhibiting factors of wound healing in diabetic patients. Plant extracts are a rich source of polyphenols, allowing them to be an effective agent for wound healing. Drying temperature and extraction solvent highly affect the stability of polyphenols in plant materials. However, there is a need to optimize the extraction protocol to ensure the efficacy of the final product. For this purpose, the effects of drying temperature and solvents on the polyphenolic composition and diabetic wound healing activity of Moringa oleifera leaves were examined in the present research. Fresh leaves were oven dried at different temperatures (10 °C, 30 °C, 50 °C, and 100 °C) and extracted in three solvents (acetone, ethanol, and methanol) to obtain twelve extracts in total. The extracts were assessed for free radical scavenging and antihyperglycemic effects using DPPH (2,2-diphenylpicrylhydrazyl) and α- glucosidase inhibition assays. Alongside this, a scratch assay was performed to evaluate the cell migration activity of M. oleifera on the human retinal pigment epithelial cell line. The cytotoxicity of the plant extracts was assessed on human retinal pigment epithelial (RPE) and hepatocellular carcinoma (Huh-7) cell lines. Using high-performance liquid chromatography, phenolic compounds in extracts of M. oleifera were identified. We found that an ethanol-based extract prepared by drying the leaves at 10 °C contained the highest amounts of identified polyphenols. Moringa oleifera extracts showed remarkable antioxidant, antidiabetic, and cell migration properties. The best results were obtained with leaves dried at 10 °C and 30 °C. Decreased activities were observed with drying temperatures of 50 °C and above. Moreover, M. oleifera extracts exhibited no toxicity on RPE cells, and the same extracts were cytotoxic for Huh-7 cells. This study revealed that M. oleifera leaves extracts can enhance wound healing in diabetic conditions due to their antihyperglycemic, antioxidant, and cell migration effects. The leaves of this plant can be an excellent therapeutic option when extracted at optimum conditions.

In vitro evaluation of immunomodulatory, anti-diabetic, and anti-cancer molecular mechanisms of Tribulus terrestris extracts.

Dampened immunity and impaired wound healing in diabetic patients may lead to diabetic foot ulcer disease, which is the leading cause of limb amputations and hospitalization. On the other hand, cancer is the most significant cause of mortality globally, accounting for over 10 million fatalities in 2020, or nearly one in every six deaths. Plants and herbs have been used to treat chronic diseases due to their essential pharmaceutical attributes, such as mitigating drug resistance, ameliorating systemic toxicities, reducing the need for synthetic chemotherapeutic agents,and strengthening the immune system. The present study has been designed to evaluate the effects of Tribulus terrestris on wound healing, cytotoxic and anti-inflammatory responses against HepG-2 liver cancer cell line. Two solvents (methanol and ethanol) were used for root extraction of T. terrestris. The wound healing potential of the extracts was studied on diabetic cell culture line by scratch assay. The anti-oxidant and cytotoxic potentials were evaluated by in vitro assays against HepG2 cell line. The methanolic root extract resulted in the coverage of robust radical scavenging or maximum inhibition of 66.72%,potent cytotoxic activity or reduced cell viability of 40.98%, and anti-diabetic activity having mighty α-glucosidase inhibition of 50.16% at a concentration of 80 µg/ml. Significant reduction in the levels of LDH leakage (56.38%), substantial ROS (48.45%) and SOD (72.13%) activities were recorededMoreover, gene expression analysis demonstrated the down-regulation of inflammatory markers (TNF-α, MMP-9, Bcl-2, and AFP) in HepG-2 cells when treated with T. terresteris methanolic extract as compared to stress. Furthermore, the down-regulation of inflammatory markers was validated through ELISA-mediated protein estimation of IL-1ß and TNF-α. It is expected that this study will lay a foundation and lead to the development of efficient but low-cost, natural herbs extract-based dressing/ointment for diabetic patients and identify potential drug metabolites to treat out-of-whack inflammatory responses involved in cancer onset, progression, and metastasis.

Vitamin C supplementation ameliorates liver function profile and antiviral treatment response in Hepatitis C patients.

An imbalance between oxidative stress and antioxidative defence mediates a variety of diseases pathogenesis. The present study aims to assess the possible outcome of supplementation of oral vitamin-C (VC), an antioxidant, in Viral Hepatitis C (HCV) treatment as an adjuvant therapy. 200 HCV-patients were selected, 100 were given Vitamin-C (1000 mg/day) along with anti HCV treatment (sofosbuvir plus daclatasvir) while the other 100 took only anti-HCV treatment for 4weeks. The serum ascorbic acid (Vitamin-C) levels and functions of the liver were tested before and after the VC supplementation. HCV patients with relatively low serum ascorbic acid showed significant improvement after the intake of vitamin C. After 4 weeks of treatment, AST, ALP, albumin, and total, direct and indirect bilirubin were improved significantly in the VC group; whereas only ALT and indirect bilirubin were improved in both groups when associated with the control subjects. Comparing the two treatment groups at 4weeks; more effective and significant improvement was observed in ALT (p<0.01), AST (p<0.001), direct (p<0.01) and indirect bilirubin (p<0.001), total proteins (p<0.001) and albumin (p<0.05) in patients with VC supplementation on anti-viral treatment compared to only anti-viral treatment group. Thus, VC supplementation improves the antiviral therapy outcome by bestowing a beneficial effect in minimizing liver damage in HCV cases.

Extracts of Eucalyptus alba Promote Diabetic Wound Healing by Inhibiting α-Glucosidase and Stimulating Cell Proliferation.

Treatment of diabetic wounds has always been a challenge for primary and acute health care. Eucalyptus alba has been reported to be used for the treatment of wounds and oxidative stress. Effects of using different temperatures and solvents for the extraction of Eucalyptus alba leaves were investigated in terms of diabetic wound healing activity. Leaves of E. alba were dried at 10°C, 30°C, 50°C, and 100°C, and dissolved in ethanol, methanol, and acetone to obtain a total of 12 extracts. All the extracts have remarkable antidiabetic, antioxidant, and cell proliferation activities. Among the tested extracts, highest activities were observed with leaves dried at 10°C and 30°C, whereas drying at 100°C resulted in the lowest activities. Ethanol-based extracts exhibited significantly increased cell proliferation compared with methanol- and acetone-based extract. The present study suggests that leaves of E. alba should be dried at temperature not more than 30°C and extracted in ethanol for optimum results. However, further studies should focus on the identification of specific bioactive compounds in E. alba leaves.

Morphological and palynological assessment of taxonomically problematic genus Paspalum based on light and scanning electron microscopy.

Present research work was carried out to clarify the variations among species of genus Paspalum morphologically and palynologically as this genus is taxonomically difficult due to having multiple similar morphologically overlapping characters which make it difficult to identify. Henceforth present research work was carried out to delimit taxa within the same genus by morphological and palynological tools through light microscopy (LM) and scanning electron microscopy (SEM). Both these tools are considered as the most useful taxonomic characters for taxonomically problematic genera. The results showed a lot of variations among morphological characters. In Paspalum dilatatum, the upper glume was ovate whereas in the other two species, the upper glume was elliptic. The upper glume apex found in P. dilatatum and Paspalum scrobiculatum was obtuse whereas in Paspalum distichum, upper glume apex was acute. Glume nerves showed variation in all three species. Paspalum distichum was 3 nerved, P. scrobiculatum was 5-7 nerved, and P. dilatatum was 5-9 nerved. All three species showed variation in lemma nerves. Paspalum scrobiculatum had 3 nerved lemma whereas in P. distichum 3-5 nerved and P. dilatatum 5-9 nerved lemma were present. In polar and equatorial view, pollen grains ranged from 25 (20-30) to 37.5 (30-45) µm. Paspalum distichum appeared to be the smallest in size whereas P. dilatatum was the largest. Exine thickness ranged from 0.75 (0.5-1) to 1.35 (1.2-1.5) µm. The higher value of pollen fertility was found in P. scrobiculatum as 87.69% and the lowest value was in P. distichum as 78.08%. Morphological keys were also given for correct identification.

Comparison of Bougainvillea spectabilis and Bougainvillea glabra species inhibited in Pakistan based on microscopic studies: Light microscope and scanning electron microscope.

The anatomical variations of two plants from the Nyctaginaceae family, Bougainvillea spectabilis and Bougainvillea glabra, were studied using light and scanning electron microscopy methods in this work. Bougainvillea is a dicotyledonous with defensive traits that can withstand extreme (hot and dry) settings; according to the findings, crystal inclusions in cells, woody spines, and an abnormal development pattern are all features that help them survive against predators and are unique to this species. The Bougainvillea plant's leaves are arranged in simple pattern, alternate to each other along stem having an undulate leaves edge and an oval form. The xylem and phloem, palisade, parenchyma midrib, spongy mesophyll, raphide crystal bundles, and trichomes were all visible when bracts and leaves were transversally sectioned and dyed with toluidine blue O (TBO). The presence of crystals was confirmed by a detailed examination of the transverse leaves by using bright-field and cross-polarizing microscopy. Dissecting microscopic examination showed that all the leaves revealed leaves venation pattern that had midvein, lateral veins areoles, and trichomes. Although trichomes have been identified on both sides, a closer look at a cleaned leaf dyed with TBO showed multicellular abundant trichomes on adaxial surface. Stomata complexes were typically found on the abaxial surface of the leaf according to epidermal peels. Present studies also showed that on adaxial side, stomata were lesser in number or were absent and also showed that the morphologies of the pavement cells on the adaxial and abaxial sides of the leaf differed.

Morphological and palynological assessment of some taxa of genus Echinochloa through light and scanning electron microscopy.

The present study was undertaken to gather information on selected species of Echinochloa based on their morphological and palyonological examination through light and scanning electron microscopy. As the Echinochloa species are most problematic grass weeds and exhibit high interspecific and intraspecific variability so due to their great morphological diversity it is difficult to recognize them. A lot of variation was observed in morphological features of studied taxa. In the present study, E. colona had more or less spreading dense racemes whereas in E. crus-galli racemes were linear to ovate. Upper glume surface was pubescent in E. colona, hispid in E. crus-galli and spinose in E. walteri. E. colona was comprised of five-nerved glumes whereas in E. crus-galli glumes were three-nerved. Although pollen basic characters in family Poaceae remains consistent but great variation was observed in pollen of studied taxa. The pollen size was quite variable among the species of genus Echinochloa. In polar view, the size of pollen grains ranged from 27.5 (20-35) µm to 40 (30-50) µm. E. crus-galli was appeared to be the smallest in size whereas E. colona was the largest.). P/E ratio ranged from 0.84 to 1.15 µm among the species. The highest value of pollen fertility was found in E. colona as 93.33% and lowest was in E. crus-galli as 87.50%. The morphological key of studied taxa is also given to study variations. In conclusion, it was observed that microscopy of morphological and palynological assessment can play a vital role in identification of taxonomically problematic taxa.

Impacts of oxalic acid-activated phosphate rock and root-induced changes on Pb bioavailability in the rhizosphere and its distribution in mung bean plant.

Rhizosphere acidification in leguminous plants can release P from the dissolution of phosphate compounds which can reduce Pb bioavailability to them via the formation of insoluble Pb compounds in their rhizosphere. A soil polluted from Pb-acid batteries effluent (SPBE), having total Pb = 639 mg kg-1, was amended with six different rates (0, 0.5, 1, 2, 4 and 6%) of oxalic acid-activated phosphate rock (OAPR) and their effects on pH, available P and bioavailable Pb concentrations in the rhizosphere and bulk soils of mung bean plant were evaluated. Furthermore, the effects of these variant OAPR rates on Pb concentrations in plant parts, bioaccumulation factor (BAF) and translocation factor (TF) for Pb in grain and traits like productivity, the activities of antioxidant enzymes, and grain biochemistry were investigated. Results revealed that increasing rates of OAPR significantly increased pH values and available P while decreased bioavailable Pb concentrations in the rhizosphere over control. The highest dissolution of P in the rhizosphere was with 4 and 6% OAPR rates. As a result, the formation of insoluble Pb compounds affected on reduced Pb concentrations in shoots, roots, and grain in addition to lower grain BAF and TF values for Pb over control. Likewise, the highest plant productivity, improved grain biochemistry, high Ca and Mg concentrations, least oxidative stress, and enhanced soil alkaline phosphatase activity were found with 4 and 6% OAPR rates. The OAPR 4% rate is suggested for reducing grain Pb concentration, cell oxidative injury, and improving grain biochemistry in mung bean.

Efficacy of chitosan-coated textile waste biochar applied to Cd-polluted soil for reducing Cd mobility in soil and its distribution in moringa (Moringa oleifera L.).

Soil pollution with Cd has promoted serious concerns for medicinal plant quality. Amending Cd-polluted soils with textile waste biochar (TWB) coated with natural polymers can lower Cd bioavailability in them and reduce associated environmental and human health risks. In this study, we explored the impacts of solely applied TWB, chitosan (CH), their mix (TWB + CH) and TWB coated with CH (TBC) in Cd-polluted soil on Cd distribution in moringa (Moringa oleifera L.) shoots and roots as well as plant-available Cd in soil. Moreover, amendments effects on plant growth, dietary quality, and antioxidative defense responses were also assessed. Results revealed that the addition of TWB, CH, and TWB + CH in Cd-polluted soil reduced Cd distribution in shoots (56%, 66%, and 63%), roots (41%, 48%, and 45%), and plant-available Cd in soil (38%, 52%, and 49%), compared to control. Interestingly, the TBC showed significantly the topmost response for reducing Cd concentrations in shoots, roots, and soil by 73%, 54%, and 58%, respectively, relative to control. Moreover, amending Cd-polluted soil with TWB, CH, and TWB + CH depicted significantly better effects on plant growth, dietary quality, and activities of soil enzymes but the topmost response was observed with TBC treatment. Compared with control, TBC improved plant growth parameters: shoot length (81%), root length (90%), shoot fresh weight (60%), root fresh weight (76%), shoot dry weight (75%), root dry weight (68%) contents of chlorophyll-a (42%) and chlorophyll-b (74%), and soil enzyme activities: urease (130%), catalase (138%), protease (71%), cellobiohydrolase (45%), acid phosphatase (34%), peroxidase (60%), ß-glucosidase (152%), chitinase (62%), and phosphomonoesterase (139%). Furthermore, TBC treatment arrested Cd-induced oxidative stress via escalating the activities of antioxidant enzymes as well as improved moringa dietary parameters (protein, tannins, lipids, alkaloids, saponins, terpenoids, flavonoids, and tocopherols contents). Such findings suggest that the TBC has an immense perspective to remediate Cd-polluted soils and prevent human health risks associated with Cd exposure through the diet.

Heterologous expression of azoreductase-encoding gene azrS of Bacillus sp. MR-1/2 for enhanced azo dye decolorization and wastewater treatment.

In Pakistan, 55% of textile exports are contributed by textile-units of Faisalabad. The effluents of these textile units, being discharged without any treatment, contain the contamination of a huge amount of synthetic azo dyes. The objective of the current research was to evaluate the contribution of an azoreductase-encoding gene (azrS) from a pre-characterized azo dye decolorizing bacterial strain Bacillus sp. MR-1/2 in a high copy number host system (pUC19-T7-Top-T) of Escherichia coli strain DH5α followed by in-silico prediction of azoreductase enzyme (AzrS) function. The recombinant cells that contained azrS had a significantly higher rate of color removal in congo red and reactive black-5 dyes when compared to wild-type MR-1/2 and E. coli DH5α after 72 h of incubation. Moreover, we were able to show that the recombinant strain significantly reduced the values of all tested parameters (pH, EC, turbidity, TSS, and COD) in actual wastewater. In support of our results, it was also predicted through bioinformatics analysis that the deduced azoreductase protein of strain MR-1/2 is linked with the dye decolorization ability of the strain through NAD(P)H-ubiquinone: oxidoreductase activity. Furthermore, we also found that the deduced protein resembled closely related proteins of protein databank in many features, yet some unique features were predicted in the enzyme activity of strain MR-1/2. It was concluded that the recombinant strain could be examined in pilot-scale experiments for textile wastewater treatment.

Associative effects of lignin-derived biochar and arbuscular mycorrhizal fungi applied to soil polluted from Pb-acid batteries effluents on barley grain safety.

While disobeying environmental regulations of Pakistan, several Pb-acid batteries recycling and repairing units discharge their effluents into water canals that irrigate arable fields. Resultantly, serious ecological risks, as well as human health hazards through consumption of edible crops grown on such Pb-polluted soils have been reported. In this experiment, we observed associative effects of amending a soil polluted from Pb-acid batteries effluents (SPB) with arbuscular mycorrhizal fungi (AMF) and lignin-derived biochar (LBC) on barley grain safety to human health. The SPB was treated with AMF inoculum (a consortium of four AMF species), lignin (LN), and LBC, as sole treatments and AMF inoculum with LN and LBC. Barley parameters involving Pb distribution in grain and other parts, grain biochemistry, and nutrition were assessed. Likewise, Pb bioavailability in SPB, AMF root colonization, soil enzymes, microbial biomass carbon (MBC), and AMF produced total glomalin related soil protein (TGSP) were also scoped. Additionally, human renal cells (HEK 293) cytotoxicity test was performed by opting barley grain-related Pb concentrations. Results show that LBC + AMF significantly reduced grain Pb concentrations below the critical limit [4.67 mg kg-1 (WHO/FAO standard)], AMF colonization, MBC, soil enzymology, and TGSP, compared to control. Likewise, rest barley parameters were also improved in this treatment. Contrary to other treatments, grain produced on LBC + AMF did not result in (a) cell apoptosis, (b) cell distortion and (c) cohesion loss. Immobilization of Pb in SPB was due to the dilution effect of Pb adsorption on LBC, AMF mycelium and TGSP which resulted in a significant drop of grain Pb concentrations below the critical limit and ultimately no harm to HEK 293 cells. Our findings endorse that grain produced at LBC + AMF treatment are safer for human consumption and will not pose health risks. The LBC + AMF application can remediate SPB for safer cereal production.

Molecular characterization of Chickpea chlorotic dwarf virus strain D in Chickpea (Cicer arietinum) from District Dera Ismail Khan Khyber Pakhtunkhwa, Pakistan.

Chickpea chlorotic dwarf virus (CpCDV), a member of genus Mastrevirus (family Geminiviridae) is an important viral pathogen of chickpea and other legume crops in Middle East, North Africa, India and Pakistan. Among sixteen known strains of CpCDV three are known to infect legume crops in Punjab province of Pakistan. In this study diversity of CpCDV was explored in Khyber Pakhtunkhwa (KP) province of Pakistan. In year 2016, during a survey in the chickpea growing areas of district Dera Ismail Khan, CpCDV infected plants were identified. Leaf samples were collected, and a diagnostic PCR confirmed mastreivrus infection in 4 out of 100 samples. From these samples full-length genome of CpCDV was amplified using specific back-to-back primers. Virus molecules were sequenced to their entirety and sequence analysis of a molecule KRF4 (GenBank accession # KY952837) showed the highest pair wise sequence identity of 97% with a CpCDV molecule (KM229787) isolated from chickpea plant. An SDT analysis revealed it to be the D strain of CpCDV and a recombination detection program (RDP) showed it to be a recombinant between C (KM229780) and L (KT634301) strains of CpCDV. Thus, further supporting the intra-species recombination for CpCDV and presence of the same strain in chickpea growing areas of Pakistan other than Punjab province. This is the first identification of CpCDV (genus Mastrevirus family Geminiviridae) from chickpea (Cicer arietinum) plants in District Dera Ismail Khan, KP province, Pakistan.

Significance of leaf morphoanatomical markers for the authentication of adultered drugs sold in herbal markets of district Lahore, Pakistan.

A number of herbal plants sold in herbal markets of Lahore are under adulteration threat which can pose harmful health effects to end-user. This adulteration problem of medicinal plants can be resolved by the implication of some valuable taxonomic parameter such as leaf epidermal anatomical characteristics. Hence, this research was aimed to provide viable anatomical markers in order to resolve this adulteration issue persisted in some common marketed medicinal plants of district Lahore, that is, Cinnamomum verum Presl., Cinnamomum tamala (Buuch.-Ham.) T.Nees&Eberm., Gymnema sylvestre (Retz.) R.Br.ex Sm., Sphaeranthus indicus Linn., Artemisia maritima Linn., Achillea millifolim L., Adhatoda vasica Nees, Butea monosperma (Lam.) Taub, and Morus nigra L. Overall multiple anatomical variations (epidermal cell shape, their length and width, type of stomata, length and width of guard cells along with presence or absence of trichomes) had been reported in the study that could be worthwhile for the correct identification of medicinal plants. Irregular shapes of epidermal cells were observed in Cinnamomum verum and Achillea millifolium while pentagonal and polygonal cells were found in their adultaerants, that is, Canella winterana and Adhatoda vasica, respectively. Types of stomata were also strikingly varied among genuine plant and its adulterant, for example, anisocytic stomata were observed in Artemisia maritima while in its adulterant (Artemisia absinthium) anomocytic stomata were found. Similarly, paracytic stomata were observed in Butea monosperma, whereas its adulterant plant (Averrhoa carambola) characteristically possessed anisocytic stomata. Hence, anatomical characteristics were proved to be a valuable taxonomic tool in resolving the adulteration issue of medicinal plants.

Microscopic techniques used for the identification of medicinal plants: A case study of Senna.

BACKGROUND: The use of quality control tool for adulteration of Senna (Cassia aungustifolia) a pharmaceutically very important. They were used for multiple health disorders such as constipation, indigestion, epilepsy, asthma, piles, migraine, and heart problems. Two different species of same family or same genus used commercially in Indo-Pak using the same medicine name Senna. One named as Senna (C. aungustifolia) and its adulterant named as Sickle Senna (Cassia obtusifolia). METHODOLOGY: These two plants were analyzed using classical microscopic techniques light microscopy and the modern chemotaxonomic traits scanning electron microscopy, fluorescence studies and phytochemical studies. RESULTS: The C. aungustifolia L. had found to be a perennial herb with trilobed pollen, diacytic, paracytic, and anisocytic stomata having smooth walled epidermal cells, whereas the C. obtusifolia stands out as a perennial shrub with spheroidal and circular pollen and paracytic type of stomata having irregular shaped epidermal cells. The powdered drug of C. aungustifolia is dark grayish green, whereas the powdered drug of C. obtusifolia is light green in color. Investigation and other techniques used in this project provided the basis for the authentication of this species.

Mitochondrial gene cytochrome c oxidase I (CO1) used for molecular identification of Bactrocera zonata in Pakistan.

Bactrocera zonata is fruit pest mostly attacked on peach and cause heavy destruction in production of peach fruits by sucking their juice. For their management, we start to detect them on basis of their molecular characterization. As mitochondrial genome encodes a gene COI used as biomarker for identification of eukaryotes including insects. In present study, we amplified COI gene and cloned into pTZ57R/T vector (Fermentas). Cloned gene was confirmed through restriction analysis and sequenced through its entirety on both strands from Macrogen (South Korea) by Sanger sequencing method. Different computational tools were utilized for comparative analysis of sequence with other related sequences retrieved from databases. Related species were identified through phylogenetic analysis using Mega 7 tool. Pairwise sequence alignment showed the sequence identity about 96% with Bactrocera zonata. By identifying the pests with more authentic molecular biomarker may help the research to control them more effectively in future.

Implication of palynological techniques for the authentication of adulterated drugs traded with the same name in different herbal markets of district Lahore, Pakistan.

Adulteration in traded medicinal plants is a significant issue nowadays and use of these adulterated medicinal plants can impose harmful impact to end user. However, this problem can be overcome by ensuring the identification of traded medicinal plants which are used in making different herbal medicines. In this regard, palynological markers are considered to be an important taxonomic tool in the identification of original medicinal plant from its adulterant. Hence this study attempted to provide particular reliable palynological markers for distinguishing selected medicinal plants from their adulterants, that is, Cinnamomum verum versus Canella winterana, Cinnamomum tamala versus Cinnamomum obtusifolium, Gymnema sylvestre versus Gymnema lactiferum, Artemisia maritima versus Artemisia absinthium, Achillea millefolium versus Adhatoda vasaka, Sphaeranthus indicus versus Sphaeranthus africanus, Averrhoa carambola versus Butea monosperma, and Morus nigra versus Morus alba. Results demonstrated great variations in multiple palynological characters between original medicinal plant and its adulterant such as in pollen size, shape, colpi length, exine, intine thickness, and fertility. In equatorial view, circular to spheroidal shape of pollen was found in A. millefolium while oblate shape was observed in A. vasaka. Similarly B. monosperma pollen was 34 µm, whereas pollen of its adulterant A. carambola was 21 µm. Moreover, colpi length of A. maritima was 11.8 µm, whereas 4.5 µm in A. absinthium. Hence it can be concluded that palynological characters are commendably helpful in identification of genuine medicinal plant from its adulterant.

Assessment of mold and yeast in some bakery products of Lahore, Pakistan based on LM and SEM.

The present investigation was designed to throw light on the microbial status of bakery products available in bakeries and supermarkets of Lahore. Different bakery samples such as biscuits, pizza, patties were collected from different localities such as Anar Kali, Chauburji, Faisal Town, Iqbal Town, Model Town, Muslim Town were investigated for mold and yeast using serial dilution technique inoculated over malt extract agar and potato dextrose agar under sterilized conditions. Isolated fungi were namely Alternaria alternata, Aspergillus niger, Aspergillus flavus, Aspergillus fumigatus, Aspergillus terreus, Curvularia americana, Fusarium solani, Penicillium digitatum, Saccharomyces cerevisiae, and Geotrichum candidum. Results depicted maximum fungal viable count in biscuits, collected from Model Town while minimum count was in the samples of Chauburji. In the case of pizza, the maximum fungal viable count was found in the sample of Muslim Town. In the case of patties, the maximum fungal viable count was found in the sample of Muslim Town while minimum count was found in the sample of Iqbal Town. Prevalence of microorganisms may be due to the use of contaminated raw material, use of polluted waters, human handling, and the use of contaminated containers. Contaminated food intake can lead to measurable signs of liver injury, inflammation, etc. Preventive measures like washing and drying of hands before preparing food, cleaning of food preparation areas, and the use of clean equipment can avoid microbes which cause hazards to human health. RESEARCH HIGHLIGHTS: Bakery products of Lahore, Pakistan is investigated for the first time on the basis of light microscopy (LM) and scanning electron microscopy (SEM) and found very significant to check the quality of bakery products or not.

Genotoxic and hematological effects of chlorpyrifos exposure on freshwater fish Labeo rohita.

Chlorpyrifos is a commonly used organophosphate insecticide that causes toxicological effects in aquatic organisms especially in fish. This study determined the effects of chlorpyrifos on the genotoxic and hematological parameters of freshwater fish, Labeo rohita. The genotoxic effects of different sublethal concentrations of chlorpyrifos were investigated in the erythrocytes of Labeo rohita (commonly known as Rohu) using the Micronucleus test. Effects of chlorpyrifos on the hematological parameters of the fish were also observed. Fish specimens were exposed to three sublethal concentrations of chlorpyrifos viz., sublethal I (SL-I, 1/6th of LC50 = ∼73.8 µg/L), sublethal II (SL-II, 1/4th of LC50 = ∼110.7 µg/L) and sublethal III (SL-III, 1/2nd of LC50 = ∼221.4 µg/L) for 96 h. Blood samples were collected at every 24 h and were subjected to the Micronucleus assay. The observed micronucleus frequencies were concentration and time-dependent. The MN induction was significantly highest (p < 0.01) at all the concentrations on 96-h exposure. During the experimental period, hematological parameters like total erythrocytes count (TEC), hemoglobin (Hb) and packed cell volume (PCV) decreased, whereas total leukocytes count (TLC) increased. This study indicated that the Micronucleus assay is a useful tool to detect genotoxic potential of chlorpyrifos in fish.